LOCUS       sequence-75849-e        2933 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Intermediate array vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2933)
  AUTHORS   Sakuma T, Ochiai H, Kaneko T, Mashimo T, Tokumasu D, Sakane Y, 
            Suzuki K, Miyamoto T, Sakamoto N, Matsuura S, Yamamoto T
  TITLE     Repeating pattern of non-RVD variations in DNA-binding modules 
            enhances TALEN activity.
  JOURNAL   Sci Rep. 2013 Nov 29;3:3379. doi: 10.1038/srep03379.
  PUBMED    24287550
REFERENCE   2  (bases 1 to 2933)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep."; 
            date: "2013-11-29"; volume: "3:3379. doi"; pages: " 
            10.1038/srep03379"
COMMENT     Sequence Label: pFUS2_b2
FEATURES             Location/Qualifiers
     source          1..2933
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     233..252
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     486..503
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(662..689)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(781..867)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     931..947
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(958..1269)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGAGLGV"
     primer_bind     1134..1150
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1160..1179
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1160..1178
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1226..1246)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1226..1244)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1265..1281)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1265..1281)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1278..1300)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1289..1305
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1313..1319,1320..1337,1338..1343))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1358..1379
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             1793..2584
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      join(2677..2933,1..332)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
       61 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
      121 acctacagcg tgagcattga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
      181 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
      241 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
      301 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
      361 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
      421 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
      481 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
      541 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
      601 gcagtgagcg caacgcaatt aatacgcgta ccgctagcca ggaagagttt gtagaaacgc
      661 aaaaaggcca tccgtcagga tggccttctg cttagtttga tgcctggcag tttatggcgg
      721 gcgtcctgcc cgccaccctc cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg
      781 atttgtccta ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc
      841 ttccgactga gcctttcgtt ttatttgatg cctggcagtt ccctactctc gcgttaacgc
      901 tagcatggat gttttcccag tcacgacgtt gtaaaacgac ggccagtctt aagcgtctca
      961 gaccccgaga ccggcgccgc tacagggcgc gtcccattcg ccattcaggc tgcgcaactg
     1021 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     1081 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     1141 gacggccagt gagcgcgcgt aatacgactc actatagggc gaattgggta ccgggccccc
     1201 cctcgaggtc ctccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat
     1261 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     1321 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     1381 ttgcgttgcg ctcactgccc gctttccacc ggtggtctca tgaccccgag acgtctagac
     1441 cagccaggac agaaatgcct cgacttcgct gctacccaag gttgccgggt gacgcacacc
     1501 gtggaaacgg atgaaggcac gaacccagtg gacataagcc tgttcggttc gtaagctgta
     1561 atgcaagtag cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg
     1621 taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttgggg tacagtctat
     1681 gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga tgttatggag
     1741 cagcaacgat gttacgcagc agggcagtcg ccctaaaaca aagttaaaca ttatgaggga
     1801 agcggtgatc gccgaagtat cgactcaact atcagaggta gttggcgtca tcgagcgcca
     1861 tctcgaaccg acgttgctgg ccgtacattt gtacggctcc gcagtggatg gcggcctgaa
     1921 gccacacagt gatattgatt tgctggttac ggtgaccgta aggcttgatg aaacaacgcg
     1981 gcgagctttg atcaacgacc ttttggaaac ttcggcttcc cctggagaga gcgagattct
     2041 ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc attccgtggc gttatccagc
     2101 taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac attcttgcag gtatcttcga
     2161 gccagccacg atcgacattg atctggctat cttgctgaca aaagcaagag aacatagcgt
     2221 tgccttggta ggtccagcgg cggaggaact ctttgatccg gttcctgaac aggatctatt
     2281 tgaggcgcta aatgaaacct taacgctatg gaactcgccg cccgactggg ctggcgatga
     2341 gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc
     2401 gccgaaggat gtcgctgccg actgggcaat ggagcgcctg ccggcccagt atcagcccgt
     2461 catacttgaa gctagacagg cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc
     2521 agatcagttg gaagaatttg tccactacgt gaaaggcgag atcaccaagg tagtcggcaa
     2581 ataaccctcg agccacccat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     2641 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     2701 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     2761 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     2821 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     2881 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agt
//
